Scintillation counting of 32P without added scintillator in aqueous solutions and organic solvents and on dry chromatographic media.
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Chromatography, Paper↗
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Automatic collector of expired 14CO2 for liquid scintillation counting.
A filtration technique was employed to trap 14CO2 continuously for liquid scintillation counting. Devices for delivering scintillator and ethanolamine solutions were combined symmetrically with two fritted-glass aspirators for alternating operation. The collector was regulated by a fraction collector timer. Trial and animal tests indicated that the described method was efficient, reliable, and more convenient for frequent collection over long periods than alternative methods. The automatic collector was used for metabolic studies of [1-14C] arachidonic acid in rats kept in metabolic cages and the results were processed by multicompartmental analysis.
Animals↗
Determination of 1,2,4-trimethylbenzene (pseudocumene) in serum of a person exposed to liquid scintillation counting solutions by GC/MS.
1,2,4-Trimethylbenzene (pseudocumene) is often used as the solvent for liquid scintillation counting solutions. In the present work, the determination of 1,2,4-trimethylbenzene in the serum of a person exposed to the solvent vapor during the handling of liquid scintillation counting cocktails is described. The identification of 1,2,4-trimethylbenzene was carried out by GC/MS. The analyte was quantified by GC with flame ionization detection. The detection limit of the method is 0.01 ppm. Two hours after exposure of a person to the solvent vapor, the concentration of 1,2,4-trimethylbenzene in her serum was found to be 0.20 ppm.
Benzene Derivatives↗
Combination of high-performance liquid chromatography and microplate scintillation counting for crop and animal metabolism studies: a comparison with classical on-line and thin-layer chromatography radioactivity detection.
Samples from crop or animal metabolism studies of pesticides were used to evaluate the performance of the combination of analytical or narrow-bore HPLC and microplate scintillation counting (TopCount). Samples with extreme matrix content such as grain and tomato extracts from crop metabolism studies as well as extracts from hen excreta or goat urine from farm animal metabolism studies could be injected, analyzed, and quantified directly without any sample pretreatment. The minimum amount of radioactivity injected was approximately 1 Bq (60 dpm). Counting times from 5 to 60 min were used for detection and quantification. These results were compared with those from classical on-line radioactivity detection and with radioactivity detection on thin-layer chromatography plates.The combination of analytical or narrow-bore HPLC and microplate scintillation counting (TopCount) offers high sensitivity and high resolution power at the same time. It could be clearly demonstrated that the combination of HPLC with microplate scintillation counting is superior to the classical on-line radioactivity detection and at least equivalent to the classical thin-layer radiochromatography regarding performance and sensitivity.
Animals↗
Scintillation counting: channels ratio and external standard channels ratio for the determination of counting efficiency in Triton X-100 based scintillants.
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Carbon Radioisotopes↗
Comparison of autoradiography, liquid scintillation counting and immunoenzymatic staining of 5-bromo-2'-deoxyuridine for measurement of unscheduled DNA synthesis and replicative DNA synthesis in rat liver.
Different methods for evaluating unscheduled DNA synthesis (UDS) and replicative DNA synthesis (RDS) were studied in hepatocytes of F344 rats exposed in vivo to dimethylnitrosamine (DMN) or CCl4. Hepatocytes were isolated and incubated in Williams' medium E supplemented with either [3H]thymidine for autoradiography or 5-bromo-2'-deoxyuridine for immunoenzymatic staining. In the method of liquid scintillation counting, the cells were incubated with [3H]thymidine with or without hydroxyurea. The nuclear fraction was isolated and the incorporation of [3H]thymidine into nuclear DNA was determined by a liquid scintillation counter. DMN at doses of 0.625-5 mg/kg body weight induced UDS of 1.6-37.9 (0 dose; -6.9) net grains/nucleus measured by autoradiography and 337-1377 (0 dose; 177) dpm/microgram DNA in the presence of hydroxyurea measured by a liquid scintillation counter. CCl4 at doses of 50-400 mg/kg body weight induced RDS in 1.5-12.1% (0 dose; 0.12%) and 1.8-14.6% (0 dose; 0.16%) of cells with the methods of autoradiography and immunoenzymatic staining, respectively, and of 2991-24256 (0 dose; 324) dpm/microgram DNA in the absence of hydroxyurea with the method of liquid scintillation counting. Similar dose-dependent induction of UDS and RDS was observed with these methods. These results suggest that the methods of liquid scintillation counting and immunoenzymatic staining have almost the same sensitivity for measuring UDS and RDS as that of autoradiography.
Animals↗
Extractive procedure for uranium determination in water samples by liquid scintillation counting.
An extractive procedure for uranium determination using liquid scintillation counting with the URAEX cocktail is described. Interference from radon and a strong influence of nitrate ion were detected in this procedure. Interference from radium, thorium and polonium emissions were very low when optimal operating conditions were reached. Quenching effects were considered and the minimum detectable activity was evaluated for different sample volumes. Isotopic analysis of samples can be performed using the proposed method. Comparisons with the results obtained with the general procedure used in alpha spectrometry with passivated implanted planar silicon detectors showed good agreement. The proposed procedure is thus suitable for uranium determination in water samples and can be considered as an alternative to the laborious conventional chemical preparations needed for alpha spectrometry methods using semiconductor detectors.
Nitrates↗
A chemiluminescent effect encountered in the liquid scintillation counting of 32P phosphate.
During the course of a kinetic enzyme study of bisphosphoglyceromutase (EC 2.7.5.4.) a chemiluminescent effect was encountered in the liquid scintillation counting of the beta-emission from 32Pi. A method is described for overcoming the effect, thereby permitting the accurate determination of the radioactive compounds. It is important to check for chemiluminescent effects early in the development of new procedures that involve liquid scintillation counting.
Hydrogen-Ion Concentration↗
The uptake and distribution of [3H]benzo[a]pyrene in the Northern pike (Esox lucius). Examination by whole-body autoradiography and scintillation counting.
The uptake and distribution of the polyaromatic hydrocarbon benzo[a]pyrene in Northern pike (Esox lucius) were investigated by whole body autoradiography and scintillation counting. [3H]Benzo[a]pyrene was administered either in the diet or in the water. The levels of this xenobiotic employed corresponded to levels found in moderately polluted water. The uptake and distribution of this compound and its metabolites were followed from 10 hr to 21 days after the initial exposure. The autoradiography patterns observed here with both routes of administration suggest, as expected, that benzo[a]pyrene is taken up through the gastrointestinal system and the gills, metabolized in the liver, and excreted in the urine and bile. Other findings indicate that the gills may not be a major route of excretion for benzo[a]pyrene and its metabolites in the Northern pike; that benzo[a]pyrene may be taken up from the water directly into the skin of this fish; that benzo[a]pyrene and its metabolites are heterogeneously distributed in the kidney of the Northern pike; and that very little radioactivity accumulates in the adipose tissue. With scintillation counting, uptake of radioactivity from the water was found to occur rapidly in all organs, reaching a plateau in most cases after about 0.8 days. The concentrations of radioactivity in different organs ranged between 50 (many organs) and 80,000 (gallbladder + bile) times that found in the surrounding water. Since most of the radioactivity recovered in different organs of the pike after 8.5 days of exposure was in the form of metabolites, we feel that metabolism may play an important role in the bioconcentration of xenobiotics in fish.
Animals↗
A new method for secondary standard measurements with the aid of liquid scintillation counting.
A new secondary standard measuring procedure based on liquid scintillation counting has been developed at PTB. In this procedure, the efficiency curve, i.e. the counting efficiency of the nuclide to be investigated as a function of the tritium counting efficiency, is-in contrast to the CIEMAT/NIST method-not calculated but determined by experiment. For the measurements, gamma-emitting activity standards are used which were calibrated by other methods. The procedure was successfully used to calibrate solutions of the radionuclides 54Mn, 57Co, 65Zn, 85Sr and 125I with small uncertainties.
Algorithms↗
234Th analysis of marine sediments via extraction chromatography and liquid scintillation counting.
234Th is widely used as a natural tracer for study of biological mixing and particle scavenging processes in the ocean. This naturally occurring nuclide serves this purpose due to its convenient half-life (24.1 days), constant rate of production from 238U dissolved in seawater, and its strong tendency to attach to particles in seawater. As a beta/gamma emitter, 234Th may be determined using low-level gas-flow proportional counting, gamma spectrometry, and liquid scintillation counting (LSC). We describe here a technique which combines Cerenkov counting to evaluate 234Th (via 234Pa) with LSC alpha counting of 230Th added to the samples as a yield tracer. Our separation approach is based on a sample preparation procedure for marine sediments using nitric acid leaching in a "hot block", and extraction chromatography (TEVA x Resin) for Th isolation. Samples are counted in plastic LSC vials, using Ultima Gold AB cocktail, in 1 M H3PO4 media. A series of sediment samples spiked with known amounts of 234Th yielded activities within a few percent of the anticipated values.
Journal Article↗
Application of simultaneous determination of 3H, 14C, and 22Na by liquid scintillation counting to the measurement of cellular ion-transport.
A liquid scintillation counting method for simultaneous determination of three radioactive nuclides (3H, 14C, and 22Na) of biological interest was studied. By comparing the beta spectra of the three nuclides, their counting energy ranges, A, B, and C, were determined. 22NA was set high enough to avoid any spillover counts from lower-energy nuclides. Region A for 3H was set to maximize the counting efficiency. A good correlation between the counting efficiency for 22Na in region C and the counting efficiency of other nuclides in all regions was obtained. Prior to 3H and 14C dpm calculations, the 22Na counts spilled down in regions A and B were subtracted from the total counts in regions A and B. A simple linear equation was then used to compute 3H and 14C dpm. Findings show that the method presented is adaptable for highly quenched samples up to quenching indices of tSIE = 100. The method is useful for studying the biological transport coupled to Na+.
Beta Particles↗
Liquid scintillation counting of aqueous samples using triton-containing scintillants.
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Benzoates↗
Sources of error in estimating radioactivity in protein from cell cultures by liquid scintillation counting.
In this study, several common sample preparation techniques for liquid scintillation counting were critically reviewed. It has been shown that techniques such as NaOH or formic acid solubilization of trichloroacetic acid (TCA)-precipitated proteins led to underestimation of the radioactivity by 20-40%; this loss was not corrected by either internal or external standardization. Hydrolysis of the proteins with 6 N HCl or Pronase significantly increased the recovery of the labeled proteins. Also, 10% of the labeled cell proteins remained on the dish when cells were scraped into buffer; these labelled proteins could be recovered either by in situ hydrolysis with Pronase or by solubilization and scraping in 0.3 N NaOH. These techniques increased the recovered radioactivity by 50-60%, allowing quantitative measurements to be made over a 3-day chase period. A possible mechanism and the implications of this observation were discussed.
Animals↗
Assessment of drinking water radioactivity content by liquid scintillation counting: set up of high sensitivity and emergency procedures.
In our institute, different procedures have been developed to measure the radioactivity content of drinking water both in normal and in emergency situations, such as those arising from accidental and terrorist events. A single radiometric technique, namely low level liquid scintillation counting (LSC), has been used. In emergency situations a gross activity screening is carried out without any sample treatment by a single and quick liquid scintillation counting. Alpha and beta activities can be measured in more than one hundred samples per day with sensitivities of a few Bq/L. Higher sensitivity gross alpha and beta, uranium and radium measurements can be performed on water samples after specific sample treatments. The sequential method proposed is designed in such a way that the same water sample can be used in all the stages, with slight modifications. This sequential procedure was applied in a survey of the Lombardia district. At first tap waters of the 13 largest towns were examined, then a more detailed monitoring was carried out in the surroundings of Milano and Lodi towns. The high sensitivity method for the determination of uranium isotopes was used to check the presence of depleted uranium in Lake Garda. Reduced equipment requirements and relative readiness of radiochemical procedures make LSC an attractive technique which can also be applied by laboratories lacking specific radiochemistry facilities and experience.
Emergencies↗
Liquid scintillation counting of 3H-thymidine incorporated into rat lens DNA.
DNA synthesis in the lens has previously been localized by autoradiography following incorporation of 3H-thymidine. For the quantification of DNA synthesis in the lens, pooling of lenses and extraction of the DNA for liquid scintillation counting, has formerly been adapted. In the present investigation a method has been developed for the extraction of the unincorporated tracer from whole lenses after short time incubation in a medium containing 3H-thymidine. The 3H-thymidine incorporated into individual lenses was then detected by liquid scintillation counting after dissolution of the lenses. The sources of the variation in the method are evaluated.
Animals↗
Protein farnesyltransferase: measurement of enzymatic activity in 96-well format using TopCount microplate scintillation counting technology.
Protein farnesyltransferase (PFT) catalyzes the transfer of the farnesyl group of farnesyl pyrophosphate (FPP) to proteins ending with a carboxy-terminal CAAX motif, forming a thioether linkage to the cysteine residue of the protein. A method is described herein for measurement of PFT activity in 96-well format using TopCount microplate scintillation counting technology. This method has the advantages of requiring only a single transfer from reaction vessels or wells of a 96-well reaction plate to the filtration wells of a 96-well Packard UniFilter GF/B filtration plate following acid precipitation and of allowing liquid scintillation counting to be conducted directly in the filtration plate without the need for either removal of the filter from the plate or transfer of the filter to liquid scintillation vials prior to radioactivity assessment. Using rat brain cytosol as the source of PFT, H-ras as the source of farnesyl acceptor protein, and [1-3H]FPP as the farnesyl donor, the incorporation of radiolabeled farnesyl residues into H-ras was found to be a linear function of both the time of incubation at 37 degrees C (up to 75 min) and the concentration of rat brain cytosolic protein present during incubation (up to 40 micrograms protein), and to be dependent on the concentration of H-ras (Km = 1.1 microM) and FPP (Km = 0.6 microM) present in the incubation reaction. In the presence of 4 microM H-ras, 0.5 microM FPP, 4 mM MgCl2, and 20 microM ZnCl2, the specific activity of rat brain cytosolic PFT measured using this methodology was 0.253 +/- 0.036 (SD; n = 30) pmol H-ras farnesylated per minute of incubation at 37 degrees C per milligram cytosolic protein. The signal-to-noise ratio for H-ras farnesylation using this methodology averaged 25 relative to incubation in the absence of H-ras (background farnesylation of cytosolic proteins) and 50 relative to incubation in the absence of both H-ras and rat brain cytosol (background filter associated radioactivity). Intraassay variability averaged 4% and interassay variability averaged 9%. FPP mimetics from a variety of distinct structural classes, previously shown to act as competitive inhibitors of the FPP-utilizing enzyme, squalene synthetase (SQS), also inhibited PFT activity measured using this methodology, but exhibited approximately 300- to 500-fold specificity for inhibition of SQS relative to inhibition of PFT, when both enzymes were measured at their respective Km FPP concentrations, suggesting structural differences between the FPP binding sites of the two enzymes.(ABSTRACT TRUNCATED AT 400 WORDS)
Alkyl and Aryl Transferases↗
Use of Triton X-100 scintillant in a simple method for the simultaneous assay of 55Fe and 59Fe by liquid scintillation counting.
A method is described using Triton X-100 scintillant to simplify liquid scintillation counting of (55)Fe alone or in combination with (59)Fe. The Triton acts as an efficient solubilizing agent allowing small volumes of aqueous solutions of iron compounds in hydrochloric acid to be counted. The efficiencies achieved are comparable with those of previously described methods whilst the technique is less time-consuming and the scintillant-aqueous mixture is stable at low temperatures. Reproducible quantitative recovery of radioactivity has been obtained.
Ascorbic Acid↗